hPSC温和消化液是阿拉丁推出的一款温和高效型细胞消化液，专为多能干细胞设计。本产品为温和消化酶，可进行单细胞传代，确保在解离细胞时最大限度减少细胞损伤，提高细胞传代存活率，消化时间3-5分钟，简化了细胞传代过程。本产品无动物来源，生产过程严格遵循ISO9001体系，并符合GMP指导原则，确保高质量和安全性。
产品特性
1.温和高效，传代存活率高
2.可进行单细胞传代。
实验准备
1.包被：使用基质胶提前1小时37℃包被培养皿。
2.试剂准备：hPSC温和消化液以及配置好的培养基提前半小时从冰箱拿出，避光，平衡至室温；PBS等其他试剂提前37℃孵育。
操作方法（此次操作以10 cm为例）
1.检查细胞：显微镜下检查细胞状态，确保细胞形态为克隆状，且边缘光滑，达到适合的汇合度（约70-75%），符合传代标准；
2.清洗：将旧培养基吸出，加入10 mL PBS冲洗细胞，避免直接冲到细胞上，以防细胞被冲掉；
3.消化：加入5 mL预热的hPSC温和消化液，将细胞放入37℃培养箱中消化3-5分钟；
4.收集：显微镜下确认细胞解离成小团块，并脱离培养板，加入使用PBS轻轻吹打细胞，将细胞悬液转移到离心管中，用PBS再次冲洗培养板，确保最大限度收集细胞。
5.制备单细胞（可选）：使用移液枪吹打6-10次，确保80-90%的细胞成为单细胞。
备注：如果需要单细胞传代，可选择本步骤。
6.离心：室温，1200 rpm离心3分钟。
7.重悬：小心吸掉上清液，加入10 mL 培养基，用移液管轻轻重悬细胞4-5次，避免产生气泡。
8.接种：将即用型基质胶包被的培养皿从37℃培养箱拿出，吸掉包被液，加入10 mL的培养基，按照推荐的细胞密度将细胞悬液接种到板中，在显微镜下检查铺板密度，确保适宜。
9.摇匀：将培养皿轻轻前后、左右摇动，确保细胞均匀分布，孵育过夜后，第二天更换培养基，每24小时更换一次培养基，直到细胞准备好进行下次操作。
hPSC Gentle Dissociation Solution is a gentle and efficient cell dissociation solution launched by Aladdin, specifically designed for pluripotent stem cells. This product is a gentle digestive enzyme that enables single-cell passaging, ensuring minimal cell damage during dissociation and improving cell survival rate after passaging. The digestion time is 3–5 minutes, which simplifies the cell passaging process. This product is of animal-free origin, and its production process strictly follows the ISO9001 system and complies with GMP guidelines, ensuring high quality and safety.
Product Features
1.Gentle and efficient, with high post-passaging survival rate.
2.Enables single-cell passaging.
Experimental Preparation
1.Coating: Use Matrigel to coat the culture dish 1 hour in advance at 37°C.
2.Reagent preparation: Take the hPSC Gentle Dissociation Solution and the prepared medium out of the refrigerator half an hour in advance, protect from light, and equilibrate to room temperature; pre-incubate other reagents such as PBS at 37°C.
Operating Method (using a 10 cm dish as an example)
1. Check cells: Observe cell status under a microscope to ensure that the cell morphology shows colonies with smooth edges, and that the confluence is appropriate (approximately 70–75%), meeting the passaging criteria.
2.Washing: Aspirate the old medium, add 10 mL PBS to rinse the cells, avoiding direct streaming onto the cells to prevent detachment.
3. Digestion: Add 5 mL of pre-warmed hPSC Gentle Dissociation Solution, and place the cells in a 37°C incubator for 3–5 minutes of digestion.
4. Collection: Confirm under a microscope that the cells have dissociated into small clumps and have detached from the culture plate. Gently pipette the cells using PBS, transfer the cell suspension to a centrifuge tube, and rinse the culture plate again with PBS to maximize cell collection.
5.Preparation of single cells (optional): Pipette 6–10 times using a pipette to ensure that 80–90% of the cells become single cells.
Note: If single-cell passaging is required, this step can be selected.
6.Centrifugation: Centrifuge at room temperature, 1200 rpm for 3 minutes.
7.Resuspension: Carefully aspirate the supernatant, add 10 mL of medium, and gently resuspend the cells 4–5 times with a pipette, avoiding bubble formation.
8.Seeding: Remove the Matrigel-coated culture dish from the 37°C incubator, aspirate the coating solution, add 10 mL of medium, inoculate the cell suspension into the plate at the recommended cell density, and check the seeding density under a microscope to ensure it is appropriate.
9.Mixing: Gently rock the culture dish back and forth and left to right to ensure even cell distribution. After overnight incubation, change the medium the next day, and replace the medium every 24 hours until the cells are ready for the next operation.
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